Laboratory of Cellular and Molecular Biology, Microbiology Team, Faculty of Biological Sciences, University of Sciences and Technology of Houari Boumediene (USTHB), PO Box 32, El Alia, Bab Ezzouar, 16111 Algiers, Algeria; Laboratory of Microbial Biotechnology and Engineering Enzymes (LMBEE), Centre of Biotechnology of Sfax (CBS), University of Sfax, Road of Sidi Mansour Km 6, PO Box 1177, Sfax 3018, Tunisia.
Laboratory of Microbial Biotechnology and Engineering Enzymes (LMBEE), Centre of Biotechnology of Sfax (CBS), University of Sfax, Road of Sidi Mansour Km 6, PO Box 1177, Sfax 3018, Tunisia.
Int J Biol Macromol. 2018 Jan;106:636-646. doi: 10.1016/j.ijbiomac.2017.08.061. Epub 2017 Aug 13.
Two extracellular peroxidases from Bjerkandera adusta strain CX-9, namely a lignin peroxidase (called LiP BA45) and manganese peroxidase (called MnP BA30), were purified simultaneously by applying successively, ammonium sulfate precipitation-dialysis, Mono-S Sepharose anion-exchange and Sephacryl S-200 gel filtration and biochemically characterized. The sequence of their NH-terminal amino acid residues showed high homology with those of fungi peroxidases. Matrix assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF/MS) analysis revealed that the purified enzymes MnP BA30 and LiP BA45 were a monomers with a molecular masses 30125.16 and 45221.10Da, respectively. While MnP BA30 was optimally active at pH 3 and 70°C, LiP BA45 showed optimum activity at pH 4 and 50°C. The two enzymes were inhibited by sodium azide and potassium cyanide, suggesting the presence of heme-components in their tertiary structures. The K and V for LiP BA45 toward 2,4-Dichlorolphenol (2,4-DCP) were 0.099mM and 9.12U/mg, respectively and for MnP BA30 toward 2,6-Dimethylphenol (2,6-DMP), they were 0.151mM and 18.60U/mg, respectively. Interestingly, MnP BA30 and LiP BA45 demonstrated higher catalytic efficiency than that of other tested peroxidases (MnP, LiP, HaP4, and LiP-SN) and marked organic solvent-stability and dye-decolorization efficiency. Data suggest that these peroxidases may be considered as potential candidates for future applications in distaining synthetic-dyes.
两株黄孢原毛平革菌（Bjerkandera adusta）菌株 CX-9 的胞外过氧化物酶，即木质素过氧化物酶（称为 LiP BA45）和锰过氧化物酶（称为 MnP BA30），通过依次应用硫酸铵沉淀-透析、Mono-S Sepharose 阴离子交换和 Sephacryl S-200 凝胶过滤进行了同时纯化，并进行了生化特性分析。它们的 NH2-末端氨基酸残基序列与真菌过氧化物酶具有高度同源性。基质辅助激光解吸电离飞行时间质谱（MALDI-TOF/MS）分析表明，纯化的 MnP BA30 和 LiP BA45 酶均为单体，分子量分别为 30125.16 和 45221.10Da。MnP BA30 的最适 pH 值为 3 和 70°C，LiP BA45 的最适 pH 值为 4 和 50°C。两种酶均被叠氮化钠和氰化钾抑制，表明其三级结构中存在血红素成分。LiP BA45 对 2,4-二氯苯酚（2,4-DCP）的 K 和 V 分别为 0.099mM 和 9.12U/mg，MnP BA30 对 2,6-二甲基苯酚（2,6-DMP）的 K 和 V 分别为 0.151mM 和 18.60U/mg。有趣的是，MnP BA30 和 LiP BA45 表现出比其他测试过的过氧化物酶（MnP、LiP、HaP4 和 LiP-SN）更高的催化效率，以及显著的有机溶剂稳定性和染料脱色效率。数据表明，这些过氧化物酶可能被认为是未来在去除合成染料方面的潜在候选酶。
